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The role of Sly-miR171d on tomato fruit chilling injury (CI) was investigated. The results showed that silencing the endogenous Sly-miR171d effectively delayed the increase of CI and electrolyte leakage (EL) in tomato fruit, and maintained fruit firmness and quality. After low temperature storage, the expression of target gene GRAS24 increased in STTM-miR171d tomato fruit, the level of GA3 anabolism and the expression of CBF1, an important regulator of cold resistance, both increased in STTM-miR171d tomato fruit, indicated that silencing the Sly-miR171d can improve the resistance ability of postharvest tomato fruit to chilling tolerance.

The miRNAs and their targets involved in senescence of strawberry fruit (Fragaria ananassa L. cv. Zhangji) were analyzed in the present study. In the previous work, three members of miR164 family, mdmmiR164d_ 1ss21AC, mdm-miR164e and mdm-miR164f_1ss21TA, and three of their targets, NAC domain transcriptional regulator superfamily protein, NAC domain containing protein 38 and NAC domain containing protein 87 had been identified by high-throughput sequencing and degradome analysis. In the process of fruit senescence from 0 to 48 h at 4°C storage, the relative levels of mdm-miR164e and mdmmiR164d_1ss21AC expression were significantly increased resulting in decreased expression of NAC genes, and delayed senescence of strawberry fruits. These results suggested that miR164 was involved in strawberry fruit senescence by negatively mediating the expression of NAC transcription factors.

One novel transcription factor gene DnWRKY29 was isolated from Dendrobium officinale, and its functions were identified by its overexpression in tobacco. Compared to the wild type (WT), the DnWRKY29 transgenic tobacco seeds and seedlings showed more sensitivity to salt and drought stresses. The seed germination rate, seedling root length, and fresh weight of transgenic tobacco were lower than in WT. The activities of peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) in transgenic tobacco were decreased, and the content of malondialdehyde (MDA) in transgenic tobacco was increased. Moreover, the expression of DnWRKY29 was up-regulated in Dendrobium seedlings under salt and drought stresses, which was consistent with the function identified by its overexpression in tobacco, indicating that the up-regulation or overexpression of DnWRKY29 impaired plant tolerance to salt and drought stresses. These results provided a new insight to the low tolerance of Dendrobium seedlings to environmental change.

Dendrobium seedlings showed low survival rate when they were transferred from in vitro conditions to greenhouse or field environment. One of the major reasons is their low tolerance to environmental changes. WRKY transcription factors are one of the largest families of transcriptional regulators in plants. They are involved in various biotic and abiotic stress responses. One DnWRKY11 gene was isolated from Dendrobium nobile. To explore the function of DnWRKY11 in Dendrobium defense responses to abiotic stress, it was overexpressed in tobacco. Under salt and drought stresses, the DnWRKY11 transgenic tobacco showed higher germination rate, longer root length, higher fresh weight, higher activities of catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), and lower content of malonidialdehyde (MDA) than the wild type. These results proved the important roles of DnWRKY11 in plant response to drought and salt stresses, and provided a potential gene for improving environmental stress tolerance of Dendrobium seedlings.

Proton (H+) and aluminum (Al3+) toxicities are major factors limiting crop production on acid soils. To study whether salicylic acid (SA) is functional in alleviating protein damage caused by H+ and Al3+ toxicities, an investigation of the antioxidant defense response regulated by SA was carried out on barley (Hordeum vulgare L.) seedlings under H+, Al3+, and combined stresses. It was found that the relative root elongation of seedlings, which grew in the solutions supplemented with SA, was significantly higher than that of seedlings without SA treatment after 24-h treatments with H+, Al3+, and combined stresses. The lesser amount of carbonylated proteins with molecular weights ranging from 14.4 to 97 kD, was accumulated in seedlings treated with SA than that in the seedlings without SA treatment. The higher activities of antioxidant enzymes and lesser content of MDA were observed in seedlings treated with SA compared with the seedlings without SA treatment. Moreover, the nitroblue tetrazolium staining of roots showed that ROS accumulation was decreased by SA treatments. This study suggested that SA could alleviate cell damage caused by H+ and Al3+ toxicities on acid soils by both activating antioxidant defense responses and reducing the contents of carbonylated proteins caused by ROS in barley seedlings.

The genome of Arabidopsis thaliana contains seven Hsp90 family genes. Three organellar and two cytosolic AtHsp90 isoforms were characterized by functionally expressing them in a temperature-sensitive Hsp90 mutant and a conditional Hsp90-null mutant of Saccharomyces cerevisiae. The cytosolic AtHsp90-1 and AtHsp90-2 showed function similar to that of yeast in chaperoning roles; they could support the growth of yeast mutants at both permissive and non-permissive temperature. Neither the full-length nor mature forms of chloroplast-located AtHsp90-5, mitochondria-located AtHsp90-6 and endoplasmic reticulum (ER)-located AtHsp90-7 could complement the yeast Hsp90 proteins. The cytosolic AtHsp90s could stabilize the biomembrane of the temperature-sensitive Hsp90 mutant strains under stress conditions, while the organellar AtHsp90s could not protect the biomembrane of the temperature-sensitive Hsp90 mutant strains. Yeast two-hybrid results showed that either pre-protein or mature forms of organellar AtHsp90s could interact with cofactors cpHsp70, Hsp70, Hsp70t-2, Cyp40, p23 and a substrate protein of NOS, while cytosolic AtHsp90s could not interact with them. These results suggest that organellar and cytosolic AtHsp90s possibly work through different molecular mechanisms in forming chaperone complexes and performing their functional roles.

The PROMOTION OF CELL SURVIVAL 1 (PCS1) gene, encoding an aspartic protease, has an important role in determining the fate of cells in embryonic development and reproduction processes in Arabidopsis. To explore the potential function of the PCS1 gene in generating reproductive sterility, we placed the PCS1 gene under the control of an 1,869-bp nucleotide sequence from the 3′ end of the second intron (AG-I) of Arabidopsis AGAMOUS and CaMV 35S (–60) minimal promoter [AG-I-35S (–60)::PCS1], and introduced it into tobacco. RT–PCR results demonstrated that the PCS1 gene driven by AG-I-35S (–60) chimeric promoter was expressed only in anthers and carpels in the reproductive tissues of transgenic tobacco. Compared to wild-type plants, all AG-I-35S (–60) and AG-I-35S (–60)::PCS1 transgenic lines showed a normal phenotype throughout the vegetative growth phase. However, during the reproductive stage, most AG-I-35S (–60)::PCS1 transgenic plant anthers displayed delayed dehiscence, failed dehiscence, petalody and hypoplasia, and the pollen grains had different shapes and sizes with a distorted, shrunken, or collapsed morphology. Moreover, three transgenic lines, PCS1-1, PCS1-3 and PCS1-4, showed higher sterility than wild-type and AG-I-35S (–60) transgenic plants, respectively. These results showed that the construct of AG-I-35S (–60)::PCS1 was partially effective at preventing seed set and provided a novel sterility strategy.

Three AtHSP90 isoforms, cytosol-localized AtHSP90.2, chloroplast-localized AtHSP90.5, and endoplasmic reticulum (ER)-localized AtHSP90.7 genes, were constitutively overexpressed in Arabidopsis thaliana to study their functional mechanisms under oxidative stress. Overexpression of AtHSP90 genes reduced germination of transgenic seeds under oxidative stress. When exposed to 10 mM H2O2, AtHSP90 transgenic seedlings displayed lower activities of superoxide dismutase, catalase, and peroxidase; higher content of malondialdehyde; and higher levels of protein damage than detected in the wild type. This indicated that overexpression of AtHSP90.2, AtHSP90.5, and AtHSP90.7 in Arabidopsis impaired plant tolerance to oxidative stress. Moreover, overexpression of chloroplast- and ER-localized AtHSP90 resulted in lower resistance to oxidative stress than that of cytosolic AtHSP90. This suggested that HSP90.2, HSP90.5, and HSP90.7 localized in different cellular compartments were involved in different functional mechanisms during oxidative stress.

Three AtHsp90 isoforms, cytosolic AtHsp90.2, chloroplast-located AtHsp90.5, and endoplasmic reticulum (ER)-located AtHsp90.7, were characterized by constitutive overexpressing their genes in Arabidopsis thaliana. Both types of the transgenic plants overexpressing cytosolic and organellar AtHsp90s showed reduced tolerance to salt and drought stresses with lower germination rates and fresh weights, but improved tolerance to high concentration of Ca(2+) comparing with the wild type plants. Transcriptional analysis of ABA-responsive genes, RD29A, RD22 and KIN2 under salt and drought stresses, indicated that the induction expression of these genes was delayed by constitutive overexpression of cytosolic AtHsp90.2, but was hardly affected by that of organellar AtHsp90.5 and AtHsp90.7. These results implied that Arabidopsis different cellular compartments-located Hsp90s in Arabidopsis might be involved in abiotic stresses by different functional mechanisms, probably through ABA-dependent or Ca(2+) pathways, and proper homeostasis of Hsp90 was critical for cellular stress response and/or tolerance in plants.

The functions of cytosolic heat shock protein AtHsp90.3 in response to heavy metal stress were characterized by using expression of AtHsp90.3 gene in yeast and Arabidopsis thaliana. AtHsp90.3 supported the Saccharomyces cerevisiae Hsp90 knockout strain R0005 growth and maintaining cells membrane integrity under cadmium and arsenic stresses, which was compatible with the components of ScHsc82 machinery. However, constitutive overexpression of AtHsp90.3 in Arabidopsis impaired plant tolerance to Cd stress with lower germination rate and shorter root length, decreased contents of phytochelatins (PCs) and glutathione (GSH), inhibited activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), and increased content of malondialdehyde (MDA). These results suggested that proper homeostasis of Hsp90 was critical for cellular response and/or tolerance to heavy metal stress in plants.